1 GAO HanDong, International Journal of Microbiology & Advanced Immunology 2013, 1:201

International Journal of Microbiology & Advanced Immunology (IJMAI)

GC-MS Analysis of Chemical Components of Taxus Chinensis Var. Mairei
Seeds
Review Article
GAO HanDong
1
*, ZHANG YanJie
2
, LU ShunBao
2
1*
College of Forest Resources and Environment Nanjing Forestry University, Naniing 210037, P.R. China
2
College of Life Science, Key Lab of Protection and Utilization of Subtropic Plant Resources, Jiangxi Normal
University, Nanchang 330022, P.R. China
*Corresponding Author:
GAO HanDong,
College of Forest Resources and Environment Nanjing Forestry University, Naniing 210037, P.R. China
E-mail: yanjiezhang0710@126.com

Accepted: May 27, 2013
Published: May 31, 2013
Citation: GAO HanDong (2013) GC-MS Analysis of Chemical Components of Taxus chinensis var. Mairei. 1:201
Copyright: 2013 GAO HanDong. This is an open-access article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium,
provided the original author and source are credited.
Abstract
The extracts of petroleum ether, methanol, ethyl acetate and ether from Taxus chinensis var. mairei seeds were ana-
lyzed by GC-MS, and relative contents were determined using a normalized method. 56 peaks were obtained from
extracts of spermoderm and 79 peaks from endosperm of T. chinensis var. mairei. Among these peaks, 37 chemical
constituents were gained, and 24 compounds of which were identifed (64.86%). There were 35 chemical con-
stituents with content more than 1%, accounting for 94.59% of the total of all chemical constituents. 32 peaks
were gained from petroleum ether extract, and 7 peaks were identifed (21.88%) with content more than 1%. 36
peaks were gained from methanol extracts, and 14 peaks were identifed (38.89%), 13 of which with content more
than 1%, accounting for 92.86% of the total of all chemical constituents. 34 peaks were gained from ethyl acetate
extracts, while 10 peaks were identifed (29.41%) with content more than 1%. 33 peaks were gained from aether
extracts, and 11 peaks were identifed (33.33%). T. chinensis var. mairei seeds have various bio-active ingredients and
a higher value for development and utilization as a medicinal plant.
Key Words: Taxus chinensis var. mairei; seed; chemical components; GC-MS; composition analysis
GAO HanDong
2013, Volume I ssue No.2
2 GAO HanDong, International Journal of Microbiology & Advanced Immunology 2013, 1:201
Introduction
Taxus chinensis var. mairei is a rare species. It is dis-
tributed within the Yangtze River basin, the Henan
Nanling Mountains, and several mountains and valleys
in Shaanxi, Gansu, and also in the Taiwan Province in
China (Li et al., 2007). It has attracted wide attention
because of the signifcant anticancer activity of taxol
present in bark, twigs, leaves and other parts of the
species (Sonia et al., 2011, Elavarasi et al., 2012, Zhao
et al., 2007, Ozols, 1995, Shao et al., 2012, Li et al.,
2008). The species has faced huge harvesting pressure
in recent years and populations have been decimated.
Thus, researchers have been seeking ways to protect T.
chinensis v. mairei populations within a framework of
forest resource management and genetic conservation.
In addition, seeds of this species have a combination
of morpho-physiological deep dormancy with under-
developed, dormant embryos, which need grow to a
certain length before seed dormancy could be broken.
Its natural reproduction is low, resulting in an endan-
gered existing state of this species. The taxol active in-
gredient of T chinensis v. mairei. is reported to have
signifcant inhibitive effects on ovarian cancer, breast,
lung, stomach, colon, melanoma, leukemia, bladder
cancer, and central nervous system tumors (Yuan, et
a1., 2002a, 2002b, 2002c, Li, et a1., 2003, Kingston et
al., 1993, Kumaran et al., 2010). At present, the taxa-
nes and non-taxanes compounds were isolated from T.
mairei (Bergstralh et al., 2006, Yang et al., 2011, Yang
et al., 2012). The polysaccharide compounds of this
species also can improve immunity and protect the
liver and other organs (Li et al., 2007). T. chinensis v.
mairei was listed as one of Chinas class key protected
wild plants in 1999.
In this paper, extracts of petroleum ether, ethyl ether,
ethyl acetate and methanol of T. mairei spermoderm
and endosperm were identifed by GC-MS to provide
reference for further development and utilization of
this species.
Materials And Methods
Experimental Materials
T. chinensis v. mairei fruits consist of scarlet or green
cuplike arils. Fruits were collected from 20-40 years
old trees in an mixed broad-leaved forest in valleys and
slopes at 400-500m above sea level in 2011, Xiushui
County Jiujiang City, Jiangxi Province. Arils and empty
seeds were foated off after fruits were collected and
macerated in water. The natural dried seeds were sealed
into polyethylene bags and stored in the refrigerator
(4C). The TGW of seeds was 65.048g
Experimental Methods
Sample Extraction and Separation
Spermoderm and endosperm (including embryos)
of T. chinensis v. mairei seeds were separated and
weighed. Spermoderm and endosperm (1200g) were
separately placed into two 1000 ml beaker after they
were grind with a mill. Afterwards the beakers were
flled with methanol (80%) and closed for extraction
at 0 to 4 C. The extract was then fltered by Bchner
funnel at intervals of 24h. The collected fltrate was
re-extracted with 80% methanol. The process was re-
peated several times until the extract turned pale, and
all extracts were mixed. The methanol extracts of T.
chinensis v. mairei spermoderm and endosperm were
crudely isolated with system solvent (Fig.1, Zu et al.
2010). The crude extracts were separated into four
groups: petroleum ether phase, ether phase, ethyl ac-
etate phase and methanol phase. And then, these or-
ganic phases of spermoderm and endosperm were
placed in a Rotary Evaporator RE-3000 concentrates
to evaporate. Finally, the organic phases were concen-
trated to 200ml and placed at 5 C.
Figure 1 The isolating process of methanol extracts with different solvents of Taxus chinensis var. mairei spermoderm and endosperm
samples
extract by
methanol
Filter for 3
Times (72h)
filtrate
residua
reduced pressure
and concentrate
(30C)
extracted with
petroleum ether
petroleum
ether phase
aqueous
phase
aqueous
phase
ether
phase
aqueous
phase
ethyl
acetate
phase
reduced
pressure and
concentrate
(60C)
methanol
phase
residua and
aqueous
phase
3 GAO HanDong, International Journal of Microbiology & Advanced Immunology 2013, 1:201
Identifcation of extractions of T. chinensis v.
mairei spermoderm and endosperm
Each above 100 ml concentrated extract was collected
and evaporated to be concentrated dry in vacuum on
a rotary evaporator. These dry matters were respec-
tively washed by the same organic solvent to obtain
3ml samples. The samples were identifed by GC-MS
in the Forest Products Chemistry, Analysis and Testing
Center of Chinese Academy of Forestry.
Analysis was carried out on a GC-MS instrument (An-
tigone Lun 6890N/5973N, USA) equipped with a
1NNOWAX quartz capillary column ( 30 m 0. 25
mm; flm thickness 0.25 m) with 50 C- 190 C col-
umn temperature; programmed temperature was 5C
min-1; Helium was used as a carrier gas; gasifcation
temperature was 280 C. MS conditions were: The
ionization mode was EI and the ionization energy was
70eV; source temperature was 200 C; collection of
current was 300A; emission current was 1mA; instru-
ment separation rate was 600; quality range was 10-500.
Finally, spectrums of all components were checked
by the inventory signal of computer-controlled and
matched with the standard spectrum (Zu et al., 2010).
Results
The peaks identifed by GC-MS showed that there
were a variety of organic compounds in T. chinensis
v. mairei spermoderm and endosperm (Fig. 2). These
compounds were mainly organic acids, benzene and es-
ter-based. 24 kinds of organic compounds were identi-
fed in T. chinensis v. mairei spermoderm. In addition,
there were 4 indefnite and 9 unknown substances in
spermoderm as shown in the table 1. Petroleum ether
phase of spermoderm included (Z) -6 - octadece-
noic acid (32.53%), unknown 1 (31.28%), erucic acid
(19.26%), and hexadecanoic acid (7.14%); ether phase
mainly included 9-18 carbon acid (43.27%), hexade-
canoic acid (15.05%), erucic acid (11.12%), oleic acid
(10.11%), ether (6.46%), octadecanoic acid (5.16%);
ethyl acetate phase of spermoderm mainly included
9 - octadecadienoic acid (44.15%), hexadecanoic acid
(35.85%), octadecanoic acid (8.06%), 3 - phenyl -2 -
acrylic acid (6.60%); methanol phase of spermoderm
mainly included erucic acid (65.09%) 9 - 18 carbon
acid (7.83%), unknown 4 (4.03%), oil acid (3.79%), and
so on.
37 organic compounds were identifed in T. chinen-
sis v. mairei endosperm, which included 5 types un-
known chemicals. Endosperm petroleum ether phase
mainly included organic compounds, (E) -9 - octade-
cenoic acid (15.32%), unknown 5 (12.31%), unknown
6 (6.69%), unknown 7 (14.10%), 9 - octadecadienoic
acid (6.40%), erucic acid (6.38%); ether phase main-
ly included erucic acid (72.25%), oleic acid (12.42%),
TimeMin
TimeMin
TimeMin
TimeMin
Methanol extraction
Petroleum extraction Petroleum
Methanol extraction Methanol extraction
4 GAO HanDong, International Journal of Microbiology & Advanced Immunology 2013, 1:201
Ethyl acetate extraction
TimeMin TimeMin
TimeMin TimeMin
Figure 2: The ion fow chart of extracts with different solvents of Taxus chinensis var . mairei seeds.
No Compositions
Molecular
formula
Contents %
petroleum ether extract methanolic extract ethyl acetate extract ether extract
endosperm spermoderm endosperm spermoderm endosperm spermoderm endosperm spermoderm
1 (2,3)- dihydro -1,1,3-trime-
thyl -3- phenyl -1H-indene
C
18
H
2
0 22.42
2 (E)-9- octadecenoic acid C
18
H
34
O
2
15.32 3.19
3 (Z)-6- octadecenoic acid C
18
H
34
O
2
32.53
4 (Z,Z)-9,12-octadecadien-
oic acid
C
18
H
32
O
2
0.49
5 (Z,Z)-9,12-oleic acid C
18
H
32
O
2
9.46 2.47 45.7 3.12
6 1,2,3,4-tetramethylbenzene C
10
H
14
8.5
7 18- jecoleic acid C
19
H
36
O
2
0.74
8 1-ethide-2,3-dimethyl
benzene
C
10
H
14
0.15
9 2,4-diphenyl-4-methyl-
2(E)-pentene
C
18
H
20
5.22
10 3- phenyl -2-acrylic acid C
9
H
8
O
2
10.48 6.6 2.21 0.97
11 9- octadecenoic acid C
18
H
34
O
2
6.4 4.36 7.83 44.15 45.45
12 phenylformic acid C
7
H
6
O
2
1.3 1.62
13 cluytyl alcohol; octacosyl
alcohol
C
28
H
58
O 1.27
14 eicosanoic acid C
20
H
40
O
2
10.31 2.1
15 caproic acid C
6
H
12
O
2
1.74
16 erucic acid C
22
H
42
O
2
6.38 65.09 7.211 72.25 11.12
17 diethyl phthalate C
12
H
14
O
4
38.79
18 pelargonic acid C
9
H
18
O
2
2.43
19 stearic acid C
18
H
36
O
2
4.59 8.06 5.16
20 hexadecanoic acid C
16
H
32
O
2
3.58 7.14 0.76 4.55 35.85 0.75 15.05
21 octanoic acid C
8
H
16
O
2
3.48
22 acetic acid C
2
H
4
O
2
6.46
23 oleic acid C
18
H
34
O
2
3.79 8.13 12.42 10.11
24 n-Heptanoic acid C
7
H
14
O
2
4.03
Table.1 Relative contents in different extracts of Taxus chinensis var . mairei seeds
Ethyl acetate extraction Ethyl acetate extraction
Aether extract
Aether extract
5 GAO HanDong, International Journal of Microbiology & Advanced Immunology 2013, 1:201
cis-9 ,10-epoxyoctadecanoic acid (8.88%); ethyl ac-
etate phase mainly included (Z, Z) -9,12 - 18 oleic acid
(45.70%), 3 - phenyl -2 - acrylic acid (10.48%), 20 acid
(10.31%), oleic acid (8.13%), erucic acid (7.21%), hexa-
decanoic acid (4.55%); methanol phase mainly includ-
ed diethyl phthalate (38.79%) , (2,3) - dihydro--1,1,3
- trimethyl -3 - phenyl-1H-indene (22.42%), 1,2,3,4 -
tetracarboxylic benzene (8.50%), 2 , 4 - 2-phenyl -4 -
methyl--2 (E) - pentene (5.22%), as shown in table 1.
Conclusion And Discussion
56 peaks of T. mairei spermoderm components and
79 peaks of endosperm were obtained by GC-MS. 37
chemicals were detected by computer on-line informa-
tion retrieval, 24 of which compounds were identi-
fed (64.86%), and there were other 4 indefnite kinds
(10.81%) and 9 unknown kinds (24.32%). 35 com-
pounds with content more than 1%, accounted for
94.59% of the total extracted compounds. 32 peaks
were obtained in petroleum ether extractions, 7 of
which were identifed (21.88%), and their content were
more than 1%. 36 peaks were obtained in methanol
extractions, 14 of which were identifed (38.89%), and
13 compounds content more than 1%, accounting for
92.86% of the total of all compounds. 34 peaks of
ethyl acetate extractions were obtained and of which
10 compounds (29.41%) were identifed, 13 of which
compounds content were more than 1%, accounting
for 92.86% of the total of all compounds. 33 peaks of
ether extracts were obtained, 11 of which compounds
(33.33%) were identifed, 2 kinds of components had
indefnite substances (6%). Their relative content of
components was identifed with the peak area normali-
zation method. The results also showed more kinds of
organic compounds were extracted from T. chinensis
v. mairei in endosperm than those in spermoderm on
the same treatment condition. The same organic com-
pounds might exist in the different organic phase of
spermoderm and endosperm, such as erucic acid also
existed in petroleum ether phase, ether phase, metha-
nol phase of spermoderm, and petroleum ether phase,
ether phase and ethyl acetate phase of endosperm
phase. 16 acids existed in petroleum ether, ethyl ether
phase, ethyl acetate phase of endosperm and methanol
phase of spermoderm.
In recent years, researchers have made deep study of
T. chinensis v. mairei, but reports on artifcial propaga-
tion and cultivation of T. mairei research were rare(Shi
et al., 2010, Yu et al., 2012a, 2012b). Because research
on suitable germination conditions was rudimentary,
identifying chemical components and its effects on
seed germination is important for protection and cul-
tivation of this species. The development and utiliza-
tion of T. chinensis v. mairei were seriously affected
by its low seed germination rate(Yang et al., 2012). As
a rare plant species, T. chinensis v. mairei should be
vigorously cultivated on precondition of ensuring the
quality of medicines, so that their medicinal value can
be sustainable and developed.
Acknowledgements
The work reported here has been funded by Projects
from Jiangxi Province Natural Science Foundation of
China (20114BAB214007, 20122BAB214025).
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